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5. Amelioration of Fibrosis via S1P Inhibition Is Regulated by Inactivation of TGF-β and SPL Pathways in the Human Cornea.

Author

Nicholas, Sarah E., Basu, Sandip K., Mandal, Nawajes, and Karamichos, Dimitrios

Subjects
*TRANSFORMING growth factors-beta, *WNT signal transduction, *SPHINGOSINE kinase, *FIBROSIS, *CORNEA, *WESTERN immunoblotting
Abstract

Human corneal fibrosis can lead to opacity and ultimately partial or complete vision loss. Currently, corneal transplantation is the only treatment for severe corneal fibrosis and comes with the risk of rejection and donor shortages. Sphingolipids (SPLs) are known to modulate fibrosis in various tissues and organs, including the cornea. We previously reported that SPLs are tightly related to both, transforming growth factor beta (TGF-β) signaling and corneal fibrogenesis. The aim of this study was to investigate the effects of sphingosine-1-phosphate (S1P) and S1P inhibition on specific TGF-β and SPL family members in corneal fibrosis. Healthy human corneal fibroblasts (HCFs) were isolated and cultured in EMEM + FBS + VitC (construct medium) on 3D transwells for 4 weeks. The following treatments were prepared in a construct medium: 0.1 ng/mL TGF-β1 (β1), 1 μM sphingosine-1-phosphate (S1P), and 5 μM Sphingosine kinase inhibitor 2 (I2). Five groups were tested: (1) control (no treatment); rescue groups; (2) β1/S1P; (3) β1/I2; prevention groups; (4) S1P/β1; and (5) I2/β1. Each treatment was administered for 2 weeks with one treatment and switched to another for 2 weeks. Using Western blot analysis, the 3D constructs were examined for the expression of fibrotic markers, SPL, and TGF-β signaling pathway members. Scratch assays from 2D cultures were also utilized to evaluate cell migration We observed reduced fibrotic expression and inactivation of latent TGF-β binding proteins (LTBPs), TGF-β receptors, Suppressor of Mothers Against Decapentaplegic homologs (SMADs), and SPL signaling following treatment with I2 prevention and rescue compared to S1P prevention and rescue, respectively. Furthermore, we observed increased cell migration following stimulation with I2 prevention and rescue groups, with decreased cell migration following stimulation with S1P prevention and rescue groups after 12 h and 18 h post-scratch. We have demonstrated that I2 treatment reduced fibrosis and modulated the inactivation of LTBPs, TGF-β receptors, SPLs, and the canonical downstream SMAD pathway. Further investigations are warranted in order to fully uncover the potential of utilizing SphK I2 as a novel therapy for corneal fibrosis. [ABSTRACT FROM AUTHOR]

Published
2024
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15. Intracellular Delivery of Drugs to Macrophages

Author

Mukhopadhyay, Amitabha, Basu, Sandip K., Scheper, T., editor, Babel, W., editor, Blanch, H. W., editor, Endo, I., editor, Enfors, S. -O., editor, Fiechter, A., editor, Hoare, M., editor, Mattiasson, B., editor, Sahm, H., editor, Schügerl, K., editor, Stephanopoulos, G., editor, von Stockar, U., editor, Tsao, D. T., editor, Villadsen, J., editor, Wandrey, C., editor, Zhong, J. -J., editor, Ghose, Tarun K., editor, Ghosh, Purnendu, editor, Basu, S. K., editor, Bhardwaj, D., editor, Chauhan, V. S., editor, Dhar, N., editor, Johri, B. N., editor, Khurana, J. P., editor, Mukhopadhyay, A., editor, Padmanaban, G., editor, Sharma, A., editor, Sharma, M., editor, Swarup, R., editor, Tyagi, A. K., editor, and Virdi, J. S., editor

Published
2003
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17. Probing misconduct

Author

Basu, Sandip K., Bhargava, P. M., Sunder, R., and Desiraju, G. R.

Published
2007

30. Mycobacterium tuberculosis 6-kDa Early Secreted Antigenic Target (ESAT-6) protein downregulates Lipopolysaccharide induced c-myc expression by modulating the Extracellular Signal Regulated Kinases 1/2

Author

Mir Fayaz, Basu Sandip K, Giang Pham H, Ganguly Niladri, Siddiqui Imran, and Sharma Pawan

Subjects
Immunologic diseases. Allergy, RC581-607
Abstract

Abstract Background Mycobacterium tuberculosis (Mtb) causes death of 2–3 million people every year. The persistence of the pathogenic mycobacteria inside the macrophage occurs through modulation of host cell signaling which allows them, unlike the other non-pathogenic species, to survive inside the host. The secretory proteins of M. tuberculosis have gained attention in recent years both as vaccine candidates and diagnostic tools; they target the immune system and trigger a putatively protective response; however, they may also be involved in the clinical symptoms of the disease. Results Our studies showed that RD-1-encoded secretory protein ESAT-6 is involved in modulation of the mitogen-activated protein (MAP) kinase-signaling pathway inside the macrophage. ESAT-6 induced phosphorylation of extracellular signal-regulated kinases 1/2 (ERK1/2) in the cytoplasm but not in the nucleus, which normally is the case for MAP kinases. ESAT-6 also antagonized LPS-induced ERK1/2 phosphorylation in the nucleus. Stimulation of cells by ESAT-6 along with sodium orthovanadate (a tyrosine phosphatase inhibitor) restored phosphorylation of ERK1/2 in the nucleus, suggesting active dephosphorylation of ERK1/2 by some putative phosphatase(s) in the nucleus. Further, ESAT-6 was found to down regulate the expression of LPS-inducible gene c-myc in an ERK1/2-dependent manner. Conclusion This study showed the effect of secretory proteins of M. tuberculosis in the modulation of macrophage signaling pathways particularly ERK1/2 MAP kinase pathway. This modulation appears to be achieved by limiting the ERK1/2 activation in the nucleus which ultimately affects the macrophage gene expression. This could be a mechanism by which secretory proteins of Mtb might modulate gene expression inside the macrophages.

Published
2007
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36. Identification of senescent cell surface targetable protein DPP4

Author

Kim, Kyoung Mi, primary, Noh, Ji Heon, additional, Bodogai, Monica, additional, Martindale, Jennifer L., additional, Yang, Xiaoling, additional, Indig, Fred E., additional, Basu, Sandip K., additional, Ohnuma, Kei, additional, Morimoto, Chikao, additional, Johnson, Peter F., additional, Biragyn, Arya, additional, Abdelmohsen, Kotb, additional, and Gorospe, Myriam, additional

Published
2017
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